Minutes 20101018 TA BIOL 208 Moffatt
From SnOwy - Ed's Wiki Notebook
Contents |
Keep these concepts clear
- isoschizomer
- isocaudamer
The tough topics this week
- omit the last three questions
Inside a vector
- a promoter
- a multi-cloning site
Prokaryotic gene structure
- promoter -- transcription start
- ribosome binding site
- start codon (RTG)
- coding sequence
- stop codon
Vectors
- for insertion -- multiple cloning site / multi-cloning site (MCS)
Transcriptional fusion
- case: we already have a promoter already + MCS
- add RBS+ATG+Gene -- variable product
- case: we already have a RBS+ATG+Gene
- add promoter -- variable activity level
- do not have to worry about whether or not this is in frame -- the ATG is on the gene (CDS) to be inserted
- do not discuss: the RBS has to be an exact distance from the ATG
Translational fusion
- case: we already have promoter+RBS+ATG+MCS
- add the Gene
- case: we already have promoter+RBS+ATG+GFP+MCS
- add the Gene -- allows for visual localization
- case: we already have promoter+RBS+ATG+His+MCS
- add the Gene -- allows us to track overexpression
- Gene (CDS) must be in frame with ATG
Do not discuss
- different variations for transcriptional fusion
Discuss
- do the flash test
- tutorial -- genomic library
